EP2211900A1 - Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique - Google Patents

Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique

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Publication number
EP2211900A1
EP2211900A1 EP08852091A EP08852091A EP2211900A1 EP 2211900 A1 EP2211900 A1 EP 2211900A1 EP 08852091 A EP08852091 A EP 08852091A EP 08852091 A EP08852091 A EP 08852091A EP 2211900 A1 EP2211900 A1 EP 2211900A1
Authority
EP
European Patent Office
Prior art keywords
calanus
composition
fish
ectoparasites
per
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP08852091A
Other languages
German (de)
English (en)
Inventor
Jan Raa
Gunnar Rørstad
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Calanus AS
Original Assignee
Calanus AS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Calanus AS filed Critical Calanus AS
Publication of EP2211900A1 publication Critical patent/EP2211900A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0003—Invertebrate antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/14—Ectoparasiticides, e.g. scabicides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/542—Mucosal route oral/gastrointestinal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine

Definitions

  • the present invention relates to novel compositions comprising copepods, such as marine Calanus species, process-for the production thereof and use of said compositions in the prevention and/or treatment of ectoparasite infestations in animals. Further the invention relates to methods for prevention and/or treatment of ectoparasite infestations in aquatic animals, particularly fish.
  • Anti-parasitic drugs such as organophosphates (trichlorvos, dichlorvos and azamethiphos) and permethrins (cypermethrin and deltamethrin) have been used to bath-treat sea-lice infested salmon.
  • Other treatments include use of chitin synthesis inhibitors such a diflubenzuron and teflubenzuron and more recently the anti- parasitic drug emamectin benzoate, administered in the feed. Due to the inevitable process of resistance development against chemically synthesized pesticides, these compounds will most probably gradually lose their antiparasitic efficacy.
  • US patent no. 5,401,727 disclose a process for stimulating the immune system of aquatic animals of the class Osteichthyes and subphylum Crustacea comprising administering an effective amount of a yeast cell wall glucan composed of glucopyranose units linked by predominantly beta- 1,3 glucosidic bonds, having at least one branch therefrom of glucopyranose units linked by beta- 1,6 glycosidic bonds. Additionally the invention provides a process for enhancing the effect of vaccines by administering an effective amount of the described yeast cell wall glucan along with vaccine antigens. Further this invention also provides a process of obtaining a glucan particularly effective for stimulating the immune system of aquatic animals of the class Osteichthyes and subphylum Crustacea.
  • beta-1, 3/1, 6-glucan When added to salmon feed, the immune modulating beta-1, 3/1, 6-glucan (MacroGard®) induces notable protection against salmon-lice infestation (Fish Farming International, July 2004, Vol. 34, No. 7, page 3), most likely by enhancing innate immune mechanisms in the skin and mucosa of the fish. Such mechanisms may include mucous immunoglobulins, lysozyme, complement factors and immune cells that guard tissue surfaces and attack intruding parasites. It is well known among those skilled in the art that beta- 1,3/1, 6-glucan triggers defence reactions by interacting with Toll-like receptors (TLRs) and with the highly specific beta- 1 ,3/1, 6-glucan receptor designated dectin.
  • TLRs Toll-like receptors
  • Beta- 1,3/1, 6-glucan is one out of many microbial structures recognized by so-called Pattern Recognition Receptors (PRRs) present in immune cells of fish and higher animals.
  • PRRs Pattern Recognition Receptors
  • the PRRs that recognize the beta- 1, 3/1, 6-glucan structure have during evolution been involved in dealing with fungal intruders and in eliciting an adequate response to such infections.
  • activation of the PRRs specific for beta- 1,3/1, 6-glucan although specifically "designed” by Nature to cope with fungal infections, also give rise to enhanced protection against virus and bacteria - and against parasites.
  • TLRs There are at least 9 different TLRs on immune cells, designed to recognize and respond to 9 different unique bacterial and viral structures (Aderem A and RJ Ulevitch. Toll-like receptors in the induction of the innate immune response. Nature 2000; 406:782-787).
  • no specific TLR for parasite structures has hitherto been found.
  • the fact that only one commercial vaccine exists against an ectoparasite is a reflection of the underlying difficulties associated with successful parasite protection by the adaptive immune system.
  • the commercial tick vaccine comprises a concealed antigen that induces the production of antibodies interfering with the ticks' ability to digest blood.
  • a corresponding strategy has been followed in Norway in research-based attempts to develop a salmon-lice vaccine. It has been demonstrated that vaccination with a concealed antigen from salmon lice induces production of antibodies that end up in the digestive organ of blood sucking salmon- lice. Challenge experiments with salmon-lice of the same type as that used in the vaccine formulation (antigens present in female salmon lice), have demonstrated that these antibodies indeed have a protective effect.
  • the present invention provides bioactive compositions comprising a copepod, processes for the production thereof, and use of said compositions for the immunization of hosts against ectoparasites which have phylogenetic and hence genetic resemblance to copepods.
  • the invention provides compositions comprising copepods of Calanus species and methods for use thereof for the immunization of aquatic animals, particularly fish, against ectoparasites.
  • a composition comprising the copepod Calanus finmarchicus and a method for the immunization of fish against sea lice such as Lepeophtheirus salmonis, Caligus elongatus and Caligus rogercresseyi is also provided.
  • the invention comprises copepod compositions that enhance innate and adaptive immune defence mechanisms involved in the rejection of parasites.
  • the invention may also be applied to prevent or treat ectoparasite infestations in host animals.
  • host animals is meant to include any fish species as well as in warm-blooded animals.
  • the sea-louse e.g. salmon louse starts its parasitic life by attaching to skin surface while it is in the so-called copepodite stage, one of the early stages in its life cycle. Of some reason it does not attach to gills, except on salmon living under experimental conditions in tank systems.
  • the salmon louse uses its rasping mouthparts as mechanical devices for penetration through the mucous and into the underlying tissues, where it finds available nutrients for growth and where it gradually develops into a blood-sucking parasite.
  • the infestation is not only the result of a mechanical rasping process, but may involve enzymatic activities that help opening up tissue structures and facilitating leakage of nutrients the parasite needs.
  • An alternative strategy for developing vaccines against ectoparasites such as salmon-lice may be to circumvent the mechanisms involved in biological adaptation between host and parasite and look for "good" antigens that are not identical to, but resemble, the parasite antigens. If antibodies against such resembling antigens also recognize antigens present in the parasite, it could be speculated that they could provide protection. Such alternative "cross-protecting" vaccines could be administered by injection, like most other vaccines are. However, since injected antigens give rise to specific antibodies primarily in the blood, the salmon-lice will not encounter these antibodies before they have developed into the blood sucking stage.
  • Calanus finmarchicus Such common structures elicit production of antibody and/or other protective principles by the fish.
  • compositions comprising the parasitic copepod and/or other non-parasitic copepods cross protection against parasitic copepods is obtained.
  • Sea lice order Siphonostomatoida
  • Lepeophtheirus and Caligus family Caligidae
  • the economically most detrimental species to fish farming are Lepeophtheirus salmonis, Caligus elongatus and Caligus rogercresseyi.
  • Atlantic salmon Salmo salar L.
  • sea-living trout are more severely affected, salmon lice have been recorded on at least 12 species in the genera Salmo (North-Atlantic salmon and trout), Salvelinus (trout and charr), and Oncorhynchus ( Pacific salmon).
  • copepods started during the geological time periods Silur-Devon and/or Jura Kritt. All copepod groups of today have developed from a common bottom living ancestor (Z. Kabata 1979. Parasitic copepoda of British Fishes. The Royal Society. British Museum (Natural History). Pp 469, ISBN 0 903874 05 9). From this ancestor the current free-living copepods have developed, including Calanus finmarchicus, and also the copepods that have developed a parasitic feeding behaviour, such as sea-lice species. Their common ancestry reveals itself in particular by the fact that that copepods belonging to various orders and genera have very similar and conserved early ontogenic stages, particularly in the so-called nauplii- and copepodite-stages.
  • Calanus is a genus of marine, free-living (planktonic) copepods found in large amounts over wide oceanic and coastal areas. In the North Atlantic, at least 4 species have been described, including C. finmarchicus, C. glacialis, C. hypeboreus and C. helgolandicus. The species Calanus finmarchicus is for instance the dominant copepod in the Nordic Seas (the Norwegian Sea and adjacent seas) both by number and biomass. C. finmarchicus is a key species constituting the main link between primary production of plant plankton and fish and mammalian species higher in the food chains in this vast area.
  • Calanus-species are naturally available as a food source to a multitude of wild fish species, including salmonids, when they are available in their nauplii- and copepodite-stages in the upper waters in spring and summer. When growing under aquaculture conditions, however, salmon, trout and other species have been deprived from this natural food organism to which they are evolutionary adapted.
  • the present invention is based on compositions comprising Calanus spp. such as C. finmarchicus, which are harvested from oceanic and coastal waters during their copepodite-stages. Moreover, the present invention is based on the fact that sea-lice of the genera Caligus and Lepeophtheirus are infecting fishes during their copepodite stages, which are phylogenetically very similar to that of Calanus. Thus, it is envisioned that a composition comprising Calanus, either alone or in combination with one or more adjuvant, may be applied to enhance the innate and adaptive mucosal immunity of fishes, so that they will enhance their ability to counteract sea-lice infestations.
  • Calanus finmarchicus may elicit innate defence as well as specific immune responses.
  • animals have developed mechanisms for early detection of chemical structures (PRRs) unique for infectious micro-organisms enabling the immune system to respond adequately and early enough to immobilize and destroy the infectious microorganism.
  • PRRs chemical structures
  • no PRRs hitherto have been described for structures present in parasites, it does not mean that such structures do not exist.
  • fish and other animals throughout evolution have developed mechanisms for early detection of, and immediate response to, intruding parasites. It is therefore an implicit part of the present invention that the bioactive effects of compositions made of Calanus species, are caused by a parasite specific PRR interacting with a signal molecule present in Calanus sp.
  • compositions comprising C ⁇ / ⁇ nws-copepodites may be in the form of unprocessed, fresh or frozen stored, or be in the form of a crude powders made by any conventional drying process, including the traditional fish-meal process. It may also include de-oiled, partially or wholly purified compositions, moist or dry, and hydrolysed C ⁇ / ⁇ wws-copepodites.
  • the compositions may either be given orally as part of feed-formulations or coated thereon, or as an immersion or bath-treatment of the fish, either alone or in combinations with one or more adjuvant which are known to enhance the innate immunity of animals, such as e.g. beta-glucans, peptidoglycans, nucleotides, oligonucleotides or mixtures thereof either synthesized or produced from yeasts, fungi and bacteria.
  • the innate immune system in which Toll-like receptors are believed to play an important role, defends the host from infestation by foreign organisms like parasites in a non-specific manner.
  • the recognition and response of the innate immune system to pathogens is generic and provide an immediate defence.
  • An overall general immune enhancement of hosts against ectoparasites which have phylogenetic and hence genetic resemblance to copepods can be obtained by injection of a purified composition comprising Calanus.
  • a general immune enhancement in fish against sea lice such as Lepeophtheirus salmonis, Caligus elongatus and Caligus rogercresseyi
  • Such compositions for administration by injection may be administered alone or optionally with one or more adjuvant and in an amount of 0.001-10 mg Calanus per kg fish, preferably 0.01-1 mg Calanus per kg fish.
  • the composition according to the invention comprises Calanus, particularly Calanus of the species Calanus finmarchicus.
  • the ectoparasites against which the immunization is directed belong to the genera Lepeophtheirus and/or Caligus, and particularly to the group Lepeophtheirus salmonis, Caligus elongatus and Caligus rogercresseyi.
  • the host animal to receive the composition is any animal exposed to ectoparasites and particularly fish, and fish particularly belonging tothe genera Salmo (North- Atlantic salmon and trout), Salvelinus (trout and charr), and/or Oncorhynchus ( Pacific salmon), and most particularly the fish is Salmo salar L.
  • a composition comprising Calanus, for the immunization of fish, particularly Salmonidae, against sea-lice.
  • the composition comprises a bioactive crude powder made by a traditional fish-meal process, or a dried or moist partially or wholly purified and de-oiled concentrate of hydrolysed Calanus.
  • the compositions may either be given orally as part of feed- formulations or coated thereon, in an amount of 0.1-100 g per kg feed, preferably 0.1-10 g per kg feed, more preferably 0.5-5 g per kg feed.
  • a method for the immunization of Salmonidae against sea lice where the Salmonidae is fed Calanus.
  • the feeding with a fodder composition comprising Calanus is commenced 0-12 weeks prior to the time when the fish is expected to become exposed to infecting sea-lice.
  • the feeding with Calanus may proceed until the earliest of the time when the fish is not longer exposed or infested.
  • a method for the immunization of Salmonidae against sea-lice where the Salmonidae is immersion treated in a bath comprising Calanus in an amount of 0.1-100 g per L, preferably 1-10 g per L, more preferably 1 -5 g per L of water for 1 -60 minutes, said water bath preferably being oxygenated and having a water temperature of 5-15 °C.
  • a composition for administration by injection is provided and administered in an amount of 0.001-10 mg Calanus per kg fish, preferably 0.01-1 mg Calanus per kg fish.
  • a method for the immunization of animals wherein a composition comprising an amount of 0.001-10 mg Calanus per kg fish, preferably 0.01-1 mg Calanus per kg fish is administered by injection.
  • a method for the immunization of Salmonidae against sea lice wherein a composition comprising Calanus, either given orally, by immersion or injected, is combined with one or more adjuvant.
  • adjuvant may be a substance known to enhance the innate or adaptive immunity of the host, such as e.g. beta-glucans, peptidoglycans, nucleotides, oligonucleotides or mixtures thereof either synthesized or produced from yeasts, fungi and bacteria.
  • the adjuvant may be given via the same or a different route than the Calanus comprising composition.
  • a composition comprising Calanus finmarchicus for the immunization of Salmonidae, particularly Atlantic salmon, against sea-lice is provided.
  • the composition comprises a bioactive crude powder made by a traditional fish-meal process, or a dried or moist partially purified and de-oiled concentrate of hydrolysed Calanus finmarchicus.
  • the compositions may either be given orally as part of feed-formulations or coated thereon, in an amount of 0.1-100 g per kg feed, preferably 0.1-10 g per kg feed, more preferably 0.5-5 g per kg feed.
  • a method for the immunization of Atlantic salmon against sea-lice where the Atlantic salmon is fed Calanus finmarchicus.
  • the feeding with a fodder composition comprising Calanus finmarchicus is commenced 0-12 weeks prior to the time when the fish is expected to become exposed to infecting sea-lice, and the feeding with said Calanus may be proceeded until the earliest of the time when the fish is no longer exposed or infested.
  • a method for the immunization of Atlantic salmon against sea-lice where the salmon is immersion treated in a bath comprising Calanus finmarchicus in an amount of 0.1- 100 g per L, preferably 1-10 g per L, more preferably 1-5 g per L of water for 1-60 minutes, said water bath preferably being oxygenated and having a water temperature of 5-15 °C.
  • a method for the immunization of Atlantic salmon against sea lice wherein a composition comprising Calanus finmarchicus, either given orally, by immersion or injection, is combined with one or more adjuvant.
  • adjuvant may be a substance known to enhance the innate or adaptive immunity of the host, such as e.g. beta-glucans, peptidoglycans, nucleotides, oligonucleotides or mixtures thereof either synthesized or produced from yeasts, fungi and bacteria.
  • the adjuvant may be given via the same or a different route than the Calanus finmarchicus comprising composition.
  • a crude powder or meal comprising C ⁇ / ⁇ nws-copepodite is manufactured from catch of Calanus by conventional methods used in the manufacturing of fish meal using suitable processing conditions.
  • the composition comprises a bioactive crude powder made by a traditional fish-meal process, or a dried or moist partially or wholly purified and de-oiled concentrate of hydrolysed Calanus.
  • the feed composition according to the present invention is manufactured by mixing or top- coating such meal of Calanus in conventional fodder compositions for fish.
  • the composition according to the present invention are suitable for use for the immunisation of a host animal against ectoparasites which are phylogenetically and immunologically similar to Calanus.
  • composition is suitable for the immunisation of fish in the genera Salmo (North- Atlantic salmon and trout), Salvelinus (trout and charr), and/or Oncorhynchus ( Pacific salmon) against ectoparasites belonging to the group Lepeophtheirus salmonis, Caligus elongatus and Caligus rogercresseyi.
  • Salmo North- Atlantic salmon and trout
  • Salvelinus trout and charr
  • Oncorhynchus Pacific salmon
  • the following non-limiting examples further illustrate certain embodiments of the invention.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Fodder In General (AREA)
  • Farming Of Fish And Shellfish (AREA)
  • Feed For Specific Animals (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne de nouvelles compositions comprenant des copépodes, tels que des espèces marines du genre Calanus, un procédé pour leur production et l'utilisation desdites compositions dans la prévention et/ou le traitement d'infestations d'animaux par des ectoparasites. En outre, l'invention concerne des procédés de prévention et/ou le traitement d'infestations par des ectoparasites d'animaux aquatiques, en particulier des poissons.
EP08852091A 2007-11-19 2008-11-17 Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique Withdrawn EP2211900A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US98882707P 2007-11-19 2007-11-19
PCT/NO2008/000409 WO2009067020A1 (fr) 2007-11-19 2008-11-17 Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique

Publications (1)

Publication Number Publication Date
EP2211900A1 true EP2211900A1 (fr) 2010-08-04

Family

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Application Number Title Priority Date Filing Date
EP08852091A Withdrawn EP2211900A1 (fr) 2007-11-19 2008-11-17 Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique

Country Status (6)

Country Link
US (1) US8057809B2 (fr)
EP (1) EP2211900A1 (fr)
CA (1) CA2705544A1 (fr)
CL (1) CL2008003427A1 (fr)
DK (1) DK200900840A (fr)
WO (1) WO2009067020A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2211900A1 (fr) * 2007-11-19 2010-08-04 Calanus AS Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique
EP2346516B1 (fr) * 2008-10-28 2015-12-23 Nofima AS Composition antimicrobienne tirée de copépodes
WO2016024168A1 (fr) 2014-08-12 2016-02-18 Institut de Recherche en Semiochimie et Ethologie Appliquee Acide palmitoléique destiné à être utilisé pour inhiber la fixation de poux du poisson sur des poissons
WO2017196607A1 (fr) * 2016-05-10 2017-11-16 Elanco Tiergesundheit Ag Composé de dihydroisoxazole à utiliser pour lutter contre les poux de mer
GB201610275D0 (en) * 2016-06-13 2016-07-27 Benchmark Animal Health Ltd Treatment of marine parasite infection

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CA2040374C (fr) * 1990-07-06 1998-06-16 Gunnar Rorstad Procede permettant d'accroitre la resistance a la maladie d'animaux aquatiques
NO300692B1 (no) * 1994-04-29 1997-07-07 Biotec Mackzymal As Solubilisert forgrenet ß-1,3-glukan og anvendelse derav samt anvendelse av usolubilisert forgrenet ß-1,3-glukan
GB9418328D0 (en) * 1994-09-12 1994-11-02 Grampian Pharm Ltd Control of sea lice in seawater fish
US6054454A (en) * 1995-12-11 2000-04-25 Novartis Corporation Method of controlling fish parasites
US6376650B1 (en) * 1998-04-16 2002-04-23 Biotec Asa Bioactive peptides, uses thereof and process for the production of same
US6538031B1 (en) * 1999-11-25 2003-03-25 Novartis Animal Health Us, Inc. Method of controlling sea lice infestation in fish
ES2575156T3 (es) 2004-07-28 2016-06-24 National Research Council Of Canada Vacunas recombinantes contra copépodos caligidae (piojo de mar) y sus secuencias antigénicas
ES2557161T3 (es) * 2005-05-26 2016-01-22 Eli Lilly And Company Producción de peces mejorada
FR2891540B1 (fr) 2005-09-30 2007-12-28 Otv Sa Procede de traitement d'eaux comprenant une etape de decantation rapide suivie d'une etape de filtration directement sur membranes de micro ou d'ultra-filtration, et dispositif correspondant.
TW200801034A (en) 2005-10-05 2008-01-01 Intervet Int Bv Novel sea lice vaccine
EP2211900A1 (fr) * 2007-11-19 2010-08-04 Calanus AS Compositions de copépode bioactives, procédés pour leur production, et utilisation de ces compositions pour prévenir ou traiter des hôtes infestés par des ectoparasites similaires au plan phylogénétique

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Also Published As

Publication number Publication date
US8057809B2 (en) 2011-11-15
CL2008003427A1 (es) 2009-06-26
US20090130149A1 (en) 2009-05-21
CA2705544A1 (fr) 2009-05-28
DK200900840A (da) 2009-07-08
WO2009067020A1 (fr) 2009-05-28
WO2009067020A4 (fr) 2009-07-23

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